Update Artikelilmiah: 25176

INEZ PALUPI

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Abstrak (Bhs. Indonesia)

Teknik rekayasa genome-editing dilakukan berbasis transformasi dengan menyisipkan konstruk plasmid CRISPR/Cas9 EgEMLP pada kalus tanaman kelapa sawit menggunakan Agrobacterium tumefaciens. Penelitian ini bertujuan untuk: 1) mengetahui pengembangan teknik delivery CRISPR/Cas9 gen EgEMLP; 2) mengetahui pengaruh konsentrasi marka seleksi higromisin terhadap pertumbuhan kalus kelapa sawit (E. guinensis); 3) mengetahui konsentrasi optimum marka seleksi higromisin pada kalus kelapa sawit transforman; 4) mengetahui perubahan basa nukleotida pada kalus kelapa sawit transforman dengan sistem CRISPR/Cas9 gen EgEMLP. Penelitian ini dilaksanakan di Pusat Penelitian Bioteknologi dan Bioindustri Indonesia, Bogor pada bulan Oktober 2018 sampai April 2019. Kegiatan penelitian dibagi menjadi 4 bagian utama yaitu, tranformasi plasmid CRISPR/Cas9 ke sel kompeten bakteri E. coli dan kalus kelapa sawit menggunakan A. tumefaciens, seleksi di media pertumbuhan, isolasi DNA genom dan sekuensing DNA. Variabel yang diamati terdiri atas kalus kelapa sawit yang hidup pada media seleksi, pita DNA hasil PCR dan susunan basa nukleotida. Hasil penelitian menunjukkan bahwa CRISPR/Cas9 gen EgEMLP telah terintergrasi ke dalam kalus kelapa sawit dilihat dari hasil validasi PCR, kemudian melalui analisis bioinformatika menggunakan software Geneious dapat terlihat ada perubahan susunan basa nitrogen pada sekuens DNA kalus transfroman.

Abtrak (Bhs. Inggris)

The CRISPR/Cas9 was transformed into the oil palm calli using Agrobacterium tumefaciens. This research was aimed to: 1) observe the development of the CRISPR/Cas9 EgEMLP delivery system; 2) determine the effect of the concentration of hygromycin as selection markers on the growth of oil palm callus (E. guinensis); 3) determine the optimum concentration of hygromycin of transformed calli; 4) determine genome-editing in nucleotide bases arrangement in CRISPR/Cas9 EgEMLP transformed plant using bioinformatics. This research was conducted at the Indonesian Research Institute of Biotechnology and Bioindustry, Bogor in October 2018 to April 2019. The research activities were divided into 4 main parts, i.e. transformation of plasmid CRISPR/Cas9 into E. coli competent cell bacteria and oil palm calli using A. tumefaciens, selection in growth media, isolation of genomic DNA and DNA sequencing. The variables measured were consisted of oil palm calli growth on selection medium, the DNA band from PCR validation and the nucleotide base arrangement from contigs. The results showed that the CRISPR/Cas9 EgEMLP was successfully integrated into the oil palm calli by observing the results of PCR validation. DNA sequence analysis indicated the presence of insertion of some bases in the target area. It was proven that the nucleotide base arrangement has edited in the transformed calli.

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